pFC83 [CMCC 3128] (ATCC® 67422)

Organism: Thermus aquaticus Brock and Freeze  /  Clone Type: Clone  /  Depositors: Cetus Corp., A Belt, Cetus Corp.

Permits and Restrictions

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Designations pFC83 [CMCC 3128]
GenBank Number

J04639

Species Thermus aquaticus Brock and Freeze
Depositors Cetus Corp., A Belt, Cetus Corp.
Vector
Construct size (kb): 7.699999809265137
Insert
DNA: genomic
Insert lengths(kb): 4.5
Gene product: Taq DNA polymerase [polI]
Insert Size (kb) 4.5
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Shipping Information

Shipped:  Freeze dried phagemid in E. coli

Comments
Restriction digests of the clone give the following sizes (kb): HindIII--4.6, 3.4; BamHI--5.1, 3.0; BglII--8.0.
Higher yields of the plasmid DNA can be obtained when the culture is grown at 30 C.
The insert contains a BamHI and a BglII site.
The sequence of the BglII/BamHI fragment is given as nucleotides -112 to +614 in the published sequence. This encodes the amino terminal portion of the protein.
A full-length Taq polymerase gene can be generated by ligating the HindIII-Asp718 fragment from pFC85 (ATCC 67421) to a 750 bp BglII-HindIII fragment from pFC83 (ATCC 67422).
References

Lawyer FC, et al. Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus. J. Biol. Chem. 264: 6427-6437, 1989. PubMed: 2649500

Gelfand DH, et al. Purified thermostable enzyme. US Patent 4,889,818 dated Dec 26 1989

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